Matrix Density Engineering of Hydrogel Nanoparticles with Simulation-Guided Synthesis for Tuning Drug Release and Cellular Uptake.

The use of a nanoparticle (NP)-based antitumor drug carrier has been an emerging strategy for selectively delivering the drugs to the tumor area and, thus, reducing the side effects that are associated with a high systemic dose of antitumor drugs. Precise control of drug loading and release is critical so as to maximize the therapeutic index of the NPs. Here, we propose a simple method of synthesizing NPs with tunable drug release while maintaining their loading ability, by varying the polymer matrix density of amine- or carboxyl-functionalized hydrogel NPs. We find that the NPs with a loose matrix released more cisplatin, with up to a 33 times faster rate. Also, carboxyl-functionalized NPs loaded more cisplatin and released it at a faster rate than amine-functionalized NPs. We performed detailed Monte Carlo computer simulations that elucidate the relation between the matrix density and drug release kinetics. We found good agreement between the simulation model and the experimental results for drug release as a function of time. Also, we compared the cellular uptake between amine-functionalized NPs and carboxyl-functionalized NPs, as a higher cellular uptake of NPs leads to improved cisplatin delivery. The amine-functionalized NPs can deliver 3.5 times more cisplatin into cells than the carboxyl-functionalized NPs. The cytotoxic efficacy of both the amine-functionalized NPs and the carboxyl-functionalized NPs showed a strong correlation with the cisplatin release profile, and the latter showed a strong correlation with the NP matrix density.

Heterogeneous Monolithic Integration of Single-Crystal Organic Materials.

Manufacturing high-performance organic electronic circuits requires the effective heterogeneous integration of different nanoscale organic materials with uniform morphology and high crystallinity in a desired arrangement. In particular, the development of high-performance organic electronic and optoelectronic devices relies on high-quality single crystals that show optimal intrinsic charge-transport properties and electrical performance. Moreover, the heterogeneous integration of organic materials on a single substrate in a monolithic way is highly demanded for the production of fundamental organic electronic components as well as complex integrated circuits. Many of the various methods that have been designed to pattern multiple heterogeneous organic materials on a substrate and the heterogeneous integration of organic single crystals with their crystal growth are described here. Critical issues that have been encountered in the development of high-performance organic integrated electronics are also addressed.

Observation of Charge Separation and Space-Charge Region in Single-Crystal P3HT/C60 Heterojunction Nanowires.

We directly observed charge separation and a space-charge region in an organic single-crystal p-n heterojunction nanowire, by means of scanning photocurrent microscopy. The axial p-n heterojunction nanowire had a well-defined planar junction, consisted of P3HT (p-type) and C60 (n-type) single crystals and was fabricated by means of the recently developed inkjet-assisted nanotransfer printing technique. The depletion region formed at the p-n junction was directly observed by exploring the spatial distribution of photogenerated carriers along the heterojunction nanowire under various applied bias voltages. Our study provides a facile approach toward the precise characterization of charge transport in organic heterojunction systems as well as the design of efficient nanoscale organic optoelectronic devices.

A non-destructive n-doping method for graphene with precise control of electronic properties via atomic layer deposition.

Graphene applications require high precision control of the Fermi level and carrier concentration via a nondestructive doping method. Here, we develop an effective n-doping technique using atomic layer deposition (ALD) of ZnO thin films on graphene through a reactive molecular layer. This ALD doping method is nondestructive, simple, and precise. The ZnO thin films on graphene are uniform, conformal, of good quality with a low density of pinholes, and finely tunable in thickness with 1 Å resolution. We demonstrate graphene transistor control in terms of the Dirac point, carrier density, and doping state as a function of the ZnO thickness. Moreover, ZnO functions as an effective thin-film barrier against air-borne water and oxygen on the graphene, resulting in extraordinary stability in air for graphene devices. ZnO ALD was also applied to other two-dimensional materials including MoS2 and WSe2, which substantially enhanced electron mobility.

Cross-stacked single-crystal organic nanowire p-n nanojunction arrays by nanotransfer printing.

We fabricated cross-stacked organic p-n nanojunction arrays made of single-crystal 6,13-bis(triisopropylsilylethynyl) pentacene (TIPS-PEN) and fullerene (C60) nanowires as p-type and n-type semiconductors, respectively, by using a nanotransfer printing technique. Single-crystal C60 nanowires were synthesized inside nanoscale channels of a mold and directly transferred onto a desired position of a flexible substrate by a lubricant liquid layer. In the consecutive printing process, single-crystal TIPS-PEN nanowires were grown in the same way and then perpendicularly aligned and placed onto the C60 nanowire arrays, resulting in a cross-stacked single-crystal organic p-n nanojunction array. The cross-stacked single-crystal TIPS-PEN/C60 nanowire p-n nanojunction devices show rectifying behavior with on/off ratio of ∼ 13 as well as photodiode characteristic with photogain of ∼ 2 under a light intensity of 12.2 mW/cm(2). Our study provides a facile, solution-processed approach to fabricate a large-area array of organic crystal nanojunction devices in a desired arrangement for future nanoscale electronics.

Single-crystal poly(3,4-ethylenedioxythiophene) nanowires with ultrahigh conductivity.

We developed single-crystal poly(3,4-ethylenedioxythiopene) (PEDOT) nanowires with ultrahigh conductivity using liquid-bridge-mediated nanotransfer printing with vapor phase polymerization. The single-crystal PEDOT nanowires are formed from 3,4-ethylenedioxythiophene (EDOT) monomers that are self-assembled and crystallized during vapor phase polymerization process within nanoscale channels of a mold having FeCl3 catalysts. These PEDOT nanowires, aligned according to the pattern in the mold, are then directly transferred to specific positions on a substrate to generate a nanowire array by a direct printing process. The PEDOT nanowires have closely packed single-crystalline structures with orthorhombic lattice units. The conductivity of the single-crystal PEDOT nanowires is an average of 7619 S/cm with the highest up to 8797 S/cm which remarkably exceeds literature values of PEDOT nanostructures/thin films. Such distinct conductivity enhancement of single-crystal PEDOT nanowires can be attributed to improved carrier mobility in PEDOT nanowires. To demonstrate usefulness of single-crystal PEDOT nanowires, we fabricated an organic nanowire field-effect transistor array contacting the ultrahigh conductive PEDOT nanowires as metal electrodes.

Nano-photosensitizers Engineered to Generate a Tunable Mix of Reactive Oxygen Species, for Optimizing Photodynamic Therapy, Using a Microfluidic Device.

This work is aimed at engineering photosensitizer embedded nanoparticles (NPs) that produce optimal amount of reactive oxygen species (ROS) for photodynamic therapy (PDT). A revised synthetic approach, coupled with improved analytical tools, resulted in more efficient PDT. Specifically, methylene blue (MB) conjugated polyacrylamide nanoparticles (PAA NPs), with a polyethylene glycol dimethacrylate (PEGDMA, Mn 550) cross-linker, were synthesized so as to improve the efficacy of cancer PDT. The long cross-linker chain, PEGDMA, increases the distance between the conjugated MB molecules so as to avoid self-quenching of the excited states or species, and also enhances the oxygen permeability of the NP matrix, when compared to the previously used shorter cross-linker. The overall ROS production from the MB-PEGDMA PAA NPs was evaluated using the traditional way of monitoring the oxidation rate kinetics of anthracence-9,10-dipropionic acid (ADPA). We also applied singlet oxygen sensor green (SOSG) so as to selectively derive the singlet oxygen (1O2) production rate. This analysis enabled us to investigate the ROS composition mix based on varied MB loading. To effectively obtain the correlation between the ROS productivity and the cell killing efficacy, a microfluidic chip device was employed to provide homogeneous light illumination from an LED for rapid PDT efficacy tests, enabling simultaneous multiple measurements while using only small amounts of NPs sample. This provided multiplexed, comprehensive PDT efficacy assays, leading to the determination of a near optimal loading of MB in a PAA matrix for high PDT efficacy by measuring the light-dose-dependent cell killing effects of the various MB-PEGDMA PAA NPs using C6 glioma cancer cells.

Modulation of hydrogel nanoparticle intracellular trafficking by multivalent surface engineering with tumor targeting peptide.

Surface engineering of a hydrogel nanoparticle (NP) with the tumor-targeting ligand, F3 peptide, enhances both the NP's binding affinity for, and internalization by, nucleolin overexpressing tumor cells. Remarkably, the F3-functionalized NPs consistently exhibited significantly lower trafficking to the degradative lysosomes than the non-functionalized NPs, in the tumor cells, after internalization. This is attributed to the non-functionalized NPs, but not the F3-functionalized NPs, being co-internalized with Lysosome-associated Membrane Protein-1 (LAMP1) from the surface of the tumor cells. Furthermore, it is shown that the intracellular trafficking of the F3-functionalized NPs differs significantly from that of the molecular F3 peptides (untethered to NPs). This has important implications for designing effective, chemically-responsive, controlled-release and multifunctional nanodrugs for multi-drug-resistant cancers.

Sonophoric nanoprobe aided pH measurement in vivo using photoacoustic spectroscopy.

Presented here is a novel method of in vivo pH sensing utilizing a hybrid optical imaging technique, photoacoustic imaging (PAI), and pH sensitive polymeric nanoprobes. Nanoprobes with hydrophobic core containing a pH sensitive dye were synthesized and used to measure the pH level ex vivo first and then in vivo by performing experiments on a rat joint model, with an achieved precision of less than 0.1 pH units. The ability of the hydrophobic functional groups in the polyacrylamide matrix to shield the molecular dye from being affected by the proteins in the plasma, and prevent the dye from leaching out, is also demonstrated.

Multifunctional nanoplatforms for fluorescence imaging and photodynamic therapy developed by post-loading photosensitizer and fluorophore to polyacrylamide nanoparticles.

We report a novel post-loading approach for constructing a multifunctional biodegradable polyacrylamide (PAA) nanoplatform for tumor-imaging (fluorescence) and photodynamic therapy (PDT). This approach provides an opportunity to post-load the imaging and therapeutic agents at desired concentrations. Among the PAA nanoparticles, a formulation containing the photosensitizer, HPPH [3-(1'-hexyloxyethyl)pyropheophorbide-a], and the cyanine dye in a ratio of 2:1 minimized the undesirable quenching of the HPPH electronic excitation energy because of energy migration within the nanoparticles and/or Förster (fluorescence) resonance energy transfer (FRET) between HPPH and cyanine dye. An excellent tumor-imaging (NIR fluorescence) and phototherapeutic efficacy of the nanoconstruct formulation is demonstrated. Under similar treatment parameters the HPPH in 1% Tween 80/5% aqueous dextrose formulation was less effective than the nanoconstruct containing HPPH and cyanine dye in a ratio of 2 to 1. This is the first example showing the use of the post-loading approach in developing a nanoconstructs for tumor-imaging and therapy.

Two-photon nano-PEBBLE sensors: subcellular pH measurements.

Intracellular pH mapping is of great importance as it plays a critical role in many cellular events. Also, in tissue, pH mapping can be an indicator for the onset of cancer. Here we describe a biocompatible, targeted, ratiometric, fluorescent, pH sensing nano-PEBBLE (Photonic Explorer for Biomedical use with Biologically Localized Embedding) that is based on two-photon excitation. Two-photon excitation minimizes the photobleaching and cell autofluorescence drastically, leading to an increase in the signal-to-noise ratio. PEBBLE nanosensors provide a novel approach for introducing membrane impermeant dyes, like HPTS, into cells. We use both non-targeted and F3 peptide targeted PEBBLE nanosensors for intracellular pH measurement of 9L cells. The intracellular measurements suggest that the non-targeted nanosensors are mostly trapped in endosomes, whereas the F3 peptide targeting enables them to escape/avoid these acidic compartments. Combining the advantages of pH sensitive PEBBLE nanoparticles, including their specific targeting, with the advantages of two-photon microscopy provides an attractive and promising prospect for non-invasive real-time monitoring of pH inside cancer cells and tissues.